• Title/Summary/Keyword: monocot

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Visualization of Multicolored in vivo Organelle Markers for Co-Localization Studies in Oryza sativa

  • Dangol, Sarmina;Singh, Raksha;Chen, Yafei;Jwa, Nam-Soo
    • Molecules and Cells
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    • v.40 no.11
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    • pp.828-836
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    • 2017
  • Eukaryotic cells consist of a complex network of thousands of proteins present in different organelles where organelle-specific cellular processes occur. Identification of the subcellular localization of a protein is important for understanding its potential biochemical functions. In the post-genomic era, localization of unknown proteins is achieved using multiple tools including a fluorescent-tagged protein approach. Several fluorescent-tagged protein organelle markers have been introduced into dicot plants, but its use is still limited in monocot plants. Here, we generated a set of multicolored organelle markers (fluorescent-tagged proteins) based on well-established targeting sequences. We used a series of pGWBs binary vectors to ameliorate localization and co-localization experiments using monocot plants. We constructed different fluorescent-tagged markers to visualize rice cell organelles, i.e., nucleus, plastids, mitochondria, peroxisomes, golgi body, endoplasmic reticulum, plasma membrane, and tonoplast, with four different fluorescent proteins (FPs) (G3GFP, mRFP, YFP, and CFP). Visualization of FP-tagged markers in their respective compartments has been reported for dicot and monocot plants. The comparative localization of the nucleus marker with a nucleus localizing sequence, and the similar, characteristic morphology of mCherry-tagged Arabidopsis organelle markers and our generated organelle markers in onion cells, provide further evidence for the correct subcellular localization of the Oryza sativa (rice) organelle marker. The set of eight different rice organelle markers with four different FPs provides a valuable resource for determining the subcellular localization of newly identified proteins, conducting co-localization assays, and generating stable transgenic localization in monocot plants.

Manipulating Isoflavone Levels in Plants

  • Jung Woo-Suk;Chung Ill-Min;Heo Hwa-Young
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.149-155
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    • 2003
  • Metabolic engineering for production of isoflavones in nonlegume plants could distribute the health benefits of these phytoestrogens in more widely-consumed grains. Series of investigation to check the ability of the heterologous isoflavone synthase enzyme to interact with the endogenous phenylpropanoid pathway have been conducted. Overall, results provide possibility of production of isoflavonoids in several plant tissue systems including soybean and nonlegumes. In tissue that undergoes naturally enhanced synthesis of anthocyanins, genistein production was enhanced. In a monocot cell system, introduced expression of a transcription factor regulating genes of the anthocyanin pathway was effective in conferring the ability to produce genistein in the presence of the isoflavone synthase gene. However, in this case the intermediate accumulated to high levels indicating an inefficiency in its conversion. Introduction of a third gene, chalcone reductase, provided the ability to synthesize an additional substrate of isoflavone synthase resulting in production of the isoflavone daidzein. These research efforts provide insight into requirements for metabolic engineering for isoflavone production in nonlegume dicot and monocot tissues.

Overcoming of Barriers to Transformation in Monocot Plants

  • Toyama Koichi;Bae, Chang-Hyu;Seo, Mi-Suk;Song, In-Ja;Lim, Yong-Pyo;Song, Pill-Soon;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.135-141
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    • 2002
  • Agrobacterium-mediated transformation has been unsuccessful for monocot plants except for a few important crops such as barley, rice, maize and wheat. We discussed here that a successful transformation of monocots demands certain critical conditions. The requirements for an efficient transformation are a selection of target tissues competent for plant regeneration and Agrobacterium-infection, and various factors promoting Agrobacterium-infection. The factors were divided into two to activate Agrobacterium and to increase plant cell's susceptibility against Agrobacterium. Optimization of these factors significantly increased transformation efficiency of zoysia grass and rice plants. A technical improvement in transformation system for monocots will promote improvement of the breed as well as a study of gene functions in monocots.

Metabolic Engineering of Isoflavone Synthesis in Soybean and Non-legumes

  • Jung, Woo-Suk
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2003.04a
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    • pp.77-84
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    • 2003
  • Metabolic engineering for production of isoflavones in non-legume plants could distribute the health benefits of these phytoe-strogens in more widely-consumed grains. We investigate the ability of the heterologous isoflavone synthase enzyme to interact with the endogenous phenylpropanoid pathway. Overall, results provide possibility of production of isoflavonoids in several plant tissue systems including soybean and non-legumes. In tissue that undergoes naturally enhanced synthesis of anthocyanins, genistein production was enhanced. In a monocot cell system, introduced expression of a transcription factor regulating genes of the antho-cyanin pathway was effective in conferring the ability produce genistein in the presence of the isoflavone synthase gene. However, in this case the intermediate accumulated to high levels indicating an inefficiency in its conversion. Introduction of a third gene, chalcone reductase, provided the ability to synthesize an additional substrate of isoflavone synthase resulting in production of the isoflavone daidzein. These research efforts provide insight into requirements for metabolic engineering for isoflavone production in non-legume dicot and monocot tissues.

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Exploring Responses to Light in the Monocot Model Plant, Brachypodium distachyon

  • Tran, Quynh-Giao;Han, Yun-Jeong;Hwang, Ok-Jin;Hoang, Quyen T.N.;Kim, Jeong-Il
    • Korean Journal of Plant Resources
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    • v.31 no.5
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    • pp.522-530
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    • 2018
  • Brachypodium distachyon has been developed as a monocot model plant for temperate grasses and bioenergy crops. Although B. distachyon research is moving forward rapidly, the study of photoresponses has not been explored. To extend our knowledge of responses to light in monocots, we performed photoresponse analysis of B. distachyon using two inbred lines, Bd21 and Bd21-3. In this study, we first compared growing phenotypes between the two lines and investigated coleoptile and primary leaf growths under dark, far-red, red, and white light conditions. The results showed that the growth of the two lines were similar until tillering stage, but other developmental stages from heading to senescence were much delayed in Bd21-3, which resulted in increased height and tiller numbers. Under different light conditions, primary leaf lengths were kept increasing during the growth period, whereas the coleoptile extension was inhibited 4 to 7 days after growth depending on the light conditions applied. These results suggest that the responses to light in B. distachyon can be examined by measuring coleoptile lengths approximately 7 days after seedling growth. Moreover, we selected light-responsive genes known in Arabidopsis thaliana, such as chlorophyll A/B binding protein (CAB), light-harvesting chlorophyll binding protein (Lhcb) and chalcone synthase (CHS), and confirmed their light-induced gene expression in B. distachyon. Therefore, the present study suggests that the inhibition of coleoptile growth can be used as the parameter to analyze photoresponses in the monocot model plant, and also provide the reference genes whose expression is induced by far-red and red light treatment.

Physiological Characterization of Mono-Cotyledonous Model Plant by Ionizing Irradiation (단자엽 모델 식물의 이온화 에너지원에 따른 생리 활성)

  • Song, Mira;Kim, Sun-Hee;Jang, Duk-Soo;Kang, Si-Yong;Kim, Jin-Baek;Kim, Sang Hoon;Ha, Bo-Keun;Park, Yong Dae;Kim, Dong Sub
    • Journal of Radiation Industry
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    • v.5 no.1
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    • pp.7-13
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    • 2011
  • The present study has been performed to compare the physiological analysis of monocot model plant (rice) in response to ionizing irradiations (cosmic-ray, gamma-ray, and Ion beam). Ionizing radiations were implanted into monocot model plant (rice) seed. After irradiation, the seeds were planted in the plastic pots for a growth period of one month. Thereafter, the morphological and physiological characteristics including malondealdehyde (MDA) and chlorophyll content, activities of antioxidant enzymes in irradiation samples were investigated. We are confirmed that the activity level of MDA and chlorophyll content were not changed by ionizing irradiation samples. However, the free radical contents were increased in all irradiated plants. And the activities of SOD, POD, and APX were significantly increased by irradiation compared with non-irradiation plant.

Molecular characterization of a novel rice(Oryza sativa L.) MAP kinase, OsEDRl, its role in defense signaling pathway.

  • Kim, Jung-A;Jwa, Nam-Soo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.82-83
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    • 2003
  • Plants have evolved differently from animals having mobile activities. Thus, plants should have developed unique defense mechanisms against biotic/abiotic stresses to which plants are differently exposed, according to seasons. Most organisms have an conserved signaling network using mitogen-activated protein kinase (MAPK) cascade(s). The phenomenon implied that they are functionally very important in all organisms. In fact, they constitute one of the major components of signaling pathways involved in regulating a wide range of cellular activities from growth and development to cell death. Recently, complete MAPK cascade was first characterized in Arabidopsis from the receptor kinase (FLS2) through fellowing MEKKI -MKK4/MKK5-MPK3/MPK6-WRKY22/MRKY29 pathway. Whereas, MAPK cascade signaling pathway in monocot plant including rice (0ryza sativa L.), the most important of all food crops and an established monocot plant research model, MAPKinase kinase kinases (MAPKKK) of rice are the first upstream component of the MAPK cascade, but MAPKKK has been first identified and characterized in our lab and designated as, OsEDRl based on its homology with the Arabidopsis EDRI. The Arabidopsis EDRl was regarded as a negative regulator of defense response and the role of rice OsEDRl was analyzed. Transcriptional regulation of OsEDRl was detected under various stresses and immunoblotting analysis is going on to detect the level of OsEDRl protein in the mutants showing unique phenotype. We also introduced the constitutively active and the dominant negative forms of the OsEDRl for characterizing biological function.

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In vivo Monitoring of the Incorporation of Chemicals into Cucumber end Rice Leaves by Chlorophyll Fluorescence Imaging

  • Kim, Jin-Hong;Jung, Ji-Eun;Lee, Choon-Hwan
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.171-178
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    • 2002
  • Chlorophyll (Chl) fluorescence imaging was used to investigate the effectiveness of in vivo incorporation methods for two chemicals, 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU) and methyl viologen (MV) in rice, a monocot, and cucumber, a dicot, leaves. four different methods (vacuum infiltration, floating, transpiration-aided incorporation through petiole and spraying) were compared, and $F_i$ and $F_v$/$F_m$ were chosen for the imaging of the DCMU- and MV-treated leaves, respectively. The effects of the chemicals in plants were generally heterogeneous over the whole leaf area. Moreover, the effectiveness of the treatment of a chemical in plant leaves was dependent on chemical species, plant species, concentration of the chemical, the treatment method, the duration of the treatment, the existence of light and detergent, etc. In conclusion, we suggest that to achieve the proposed effects of chemicals in plants for an actual experiment, these factors must be considered before the chemical treatment, and the best method for the treatment of the chemicals tested was floating and vacuum infiltration in the dicot and the monocot leaves, respectively, as drawn from Chl fluorescence imaging analysis.